| This paper as the seeds of M.sieboldii K.Koch as the research object, M.sieboldii K.Koch ’seeds r was soaked for 72h with clean water, dealing with variable-temperature stratification mixed by wet sand with 1:3. Study on the embryo development, seed anatomical structure, carbohydrate change and differentially expressed protein analysis research, knowing the relationship between carbohydrate metabolism and seed germination; to explore the localization of related protein in metabolic pathways, in order to reveal the molecular mechanisms of seed germination. Main results of this study were as follows:1. During stratification process, M.sieboldii seed soluble sugar and starch content both in the overall downward trend, α-amylase activities decline after rising first, β-amyloid activities contrary trend, two amylase activity were recovered in the late stage of seed germination. According to M. sieboldii seed germination change of carbohydrate content and amylase, seed dormancy and germination process can be divided into three stages, embryo physiology active reconstruction periods (0-45d), embryo after-ripening and dormancy-breaking periods (45-90d), embryo rapid growth periods (90d to seed germination).2. During stratification process, embryo with slowing growth periods(dry seeds to 45d), stagnant growth periods(45-75d), rapid growth periods(75d to seed germination), with "J" shaped curve.The growth stages of seed embryos were consistent with the carbohydrate changes in the process of seed stratification. The nutrients in endosperm cells which close to embryos were preferentially degraded by SEM, Disintegrated cells became the formation of large molecular nutrition transportation channel between embryo and endosperm, and complete degradated until seed germination. When the seed germination, the endosperm cells were not completely degraded, the surface of the residual starch granules was smooth, and the starch was no amylase, and the metabolism of the starch in the endosperm was terminated.3. According to the results of anatomical observation, five seeds protein samples in stratification dry seeds,0 days,45 days,90 days,110 days were separated, analyzing differential protein expression and biological function in different periods by proteomics and MS+MS/MS technology. Five samples components was detected in 2-DE mapping on average to 1462，1453、1459、1406 and 1414 repeatability protein, 72 protein points with expression changes more than 2.5 times were selected,67 differentially expressed proteins were obtained, and the success rate was 93%. Which includes 22 proteins were up-regulated, 29 proteins were down-regulated, and 16 proteins fluctuated.4. Bioinformatics analysis showed that the function of protein spots was divided into 9 groups, including:the protein synthesis, folding and stability (10%); protein translation and biosynthesis(13%); stress/defense related protein (23%);carbohydrate metabolism (10%); proteolytic related protein (12%); energy metabolism (8%); signal transduction proteins (9%); storage protein (12%); unknown protein (3%).5. According to the analysis of KEGG metabolic pathway, 8 different expression proteins related to carbon metabolism with different development stages were involved into 6 metabolism pathways. Which incude 5 kinds of proteins located in glycolysis/gluconeogenesis were up-regulated at the early stage of seed germination. And it showed that seeds provided energy for seed germination by glycolysis/gluconeogenesis at the early stage of seed germination. |
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